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. 2009 Apr 1;20(7):2030–2040. doi: 10.1091/mbc.E08-11-1149

Figure 5.

Figure 5.

Tetraspanin effects on MT1-MMP–dependent pro-MMP-2 activation. (A) MCF-7-MT1 cells were treated with control or CD9/CD81/TSPAN12 (3xTSPAN) siRNAs and after 4 d, cells were plated and incubated overnight in serum-free media supplemented with conditioned media from MCF-7 cells stably expressing MMP-2. Eighteen hours later, conditioned media and lysates were collected and analyzed for pro-MMP-2 activation by gelatin zymography or actin expression by Western blotting (as loading control), respectively. Densitometry was performed on zymograms and results displayed as percentage of active MMP-2 ± SD (n = 3; *p < 0.01 compared with control). (B) HT1080 cells were transfected with control or CD81/CD151/TSPAN12 (3xTSPAN) siRNAs. After 4 d, cells were plated and then incubated overnight in serum-free media with or without 4 μM cyto D. Conditioned media were then collected and analyzed by gelatin zymography for pro-MMP-2 activation. Zymogram densitometry was performed (n = 4; *p < 0.05 compared with control). (C) MCF-7-VC cells were treated and analyzed as described in A (n = 2).